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. 2018 Apr 10;10(4):1384–1397. doi: 10.1016/j.stemcr.2018.03.006

Figure 4.

Figure 4

PLAG1-S and USF2 Collaboratively Regulate MSI2 Promoter Activity and MSI2 Expression

(A) Reporter activity upon combinatorial binding site mutagenesis for USF2 and PLAG1 within the MSI2 minimal promoter region. n = 11 independent experiments.

(B) Reporter construct luciferase activity upon independent or coincident overexpression of USF2 and PLAG1-S (n = 3 experiments).

(C) Luciferase activity of PLAG1 and USF2 MUT reporters upon PLAG1-S and USF2 co-overexpression (n = 3 experiments).

(D and E) Endogenous MSI2 transcript (D) (n = 3 experiments) and protein levels (E) following overexpression of PLAG1-S and USF2 in K562 cells compared with vector control.

(F) Luciferase reporter activity of the MSI2 minimal promoter region upon PLAG1-S or USF2 overexpression when the binding site for the reciprocal factor or for both factors is mutated (MUT) (n = 3 experiments).

(G) Co-immunoprecipitation analysis performed on K562 cells using anti-USF2 for immunoprecipitation and immunoblotting with antibodies to USF2, the known USF2 interacting partner USF1 or PLAG1.

Data are presented as means ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. See also Table S3.